Bispecific antibodies comprising an antigen-binding site binding to lag3

Abstract

The invention relates to novel antibodies particularly suitable for cancer therapies. The antibodies according to the invention are bispecific or multispecific antibodies and comprise a first antigen binding site that binds to LAG3. The first antigen binding site is an autonomous VH domain.

1 . A bispecific or multispecific antibody comprising a first antigen binding site that binds to LAG3, wherein the first antigen binding site is an autonomous VH domain. 
     
     
         2 . A bispecific or multispecific antibody of  claim 1  comprising a second antigen-binding site that binds to PD1. 
     
     
         3 . The bispecific or multispecific antibody of  claim 1  or  2 , wherein the autonomous VH domain comprises cysteines in positions (i) 52a and 71 or (ii) 33 and 52 according to Kabat numbering, wherein said cysteines form a disulfide bond under suitable conditions. 
     
     
         4 . The bispecific or multispecific antibody of any of  claims 1  to  3 , wherein the autonomous VH domain binding to LAG3 comprises
 (i) CDR1 with the sequence of SEQ ID NO: 146, a CDR2 with the sequence of SEQ ID NO: 147 and a CDR3 with the sequence of SEQ ID NO: 148; or 
 (ii) CDR1 with the sequence of SEQ ID NO: 149, CDR2 with the sequence of SEQ ID NO: 150 and CDR3 with the sequence of SEQ ID NO: 151; or 
 (iii) CDR1 with the sequence of SEQ ID NO: 152, CDR2 with the sequence of SEQ ID NO: 153 and CDR3 with the sequence of SEQ ID NO: 154; or 
 (iv) CDR1 with the sequence of SEQ ID NO: 155, CDR2 with the sequence of SEQ ID NO: 156 and CDR3 with the sequence of SEQ ID NO: 157; or 
 (v) CDR1 with the sequence of SEQ ID NO: 158, CDR2 with the sequence of SEQ ID NO: 159 and CDR3 with the sequence of SEQ ID NO: 160; or 
 vi) CDR1 with the sequence of SEQ ID NO: 161, CDR2 with the sequence of SEQ ID NO: 162 and CDR3 with the sequence of SEQ ID NO: 163; or 
 (vii) CDR1 with the sequence of SEQ ID NO: 164, CDR2 with the sequence of SEQ ID NO: 165 and CDR3 with the sequence of SEQ ID NO: 166; or 
 (viii) CDR1 with the sequence of SEQ ID NO: 167, CDR2 with the sequence of SEQ ID NO: 168 and CDR3 with the sequence of SEQ ID NO: 169; or 
 (ix) CDR1 with the sequence of SEQ ID NO: 170, CDR2 with the sequence of SEQ ID NO: 171 and CDR3 with the sequence of SEQ ID NO: 172; or 
 (x) CDR1 with the sequence of SEQ ID NO: 173, CDR2 with the sequence of SEQ ID NO: 174 and CDR3 with the sequence of SEQ ID NO: 175; or 
 xi) CDR1 with the sequence of SEQ ID NO: 176, CDR2 with the sequence of SEQ ID NO: 177 and CDR3 with the sequence of SEQ ID NO: 178. 
 
     
     
         5 . The bispecific or multispecific antibody of any of  claims 1  to  4 , wherein the autonomous VH domain comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 77, SEQ ID NO: 79, SEQ ID NO: 81, SEQ ID NO: 83, SEQ ID NO: 85 SEQ, ID NO: 87, SEQ ID NO: 89, SEQ ID NO: 91, SEQ ID NO: 93, SEQ ID NO: 95, SEQ ID NO: 97. 
     
     
         6 . The bispecific or multispecific antibody of any of  claims 1  to  5 , wherein the autonomous VH domain further comprises a substitution selected from the group consisting of H35G, Q39R, L45E and W47L. 
     
     
         7 . The bispecific or multispecific antibody of any of  claims 1  to  6 , wherein the autonomous VH domain further comprises a substitution selected from the list consisting of L45T, K94S and L108T. 
     
     
         8 . The bispecific or multispecific antibody of any of  claims 1  to  7 , wherein the autonomous VH domain comprises a VH3_23 human framework, particularly based on the VH framework of Herceptin® (trastuzumab). 
     
     
         9 . The bispecific or multispecific antibody of any of  claims 2  to  8 , wherein said second antigen-binding site binding to PD1 comprises
 a VH domain comprising 
 (i) CDR-H1 comprising the amino acid sequence of SEQ ID NO: 201, 
 (ii) CDR-H2 comprising the amino acid sequence of SEQ ID NO: 202, and 
 (iii) CDR-H3 comprising an amino acid sequence of SEQ ID NO: 203; and 
 a VL domain comprising 
 (i) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 204; 
 (ii) CDR-L2 comprising the amino acid sequence of SEQ ID NO: 205, and 
 (iii) CDR-L3 comprising the amino acid sequence of SEQ ID NO: 206. 
 
     
     
         10 . The bispecific or multispecific antibody of any of  claims 2  to  9 , wherein said second antigen-binding site binding to PD1 comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 192 and/or a VL domain comprising the amino acid sequence of SEQ ID NO: 193. 
     
     
         11 . The bispecific or multispecific antibody of any one of  claims 1  to  10 , wherein the bispecific or multispecific antibody is a human, humanized or chimeric antibody. 
     
     
         12 . The bispecific or multispecific antibody of any one of  claims 2  to  11 , wherein the bispecific or multispecific antibody comprises an Fc domain and a Fab fragment comprising the second antigen-binding site that binds to PD1. 
     
     
         13 . The bispecific or multispecific antibody of  claim 12 , wherein the Fc domain is an IgG, particularly an IgG1 Fc domain or an IgG4 Fc domain. 
     
     
         14 . The bispecific or multispecific antibody of  claim 12  or  13 , wherein the Fc domain comprises one or more amino acid substitution that reduces binding to an Fc receptor, in particular towards Fey receptor. 
     
     
         15 . The bispecific or multispecific antibody of any one of  claims 12  to  14 , wherein the Fc domain is of human IgG1 subclass with the amino acid mutations L234A, L235A and P329G (numbering according to EU index according to Kabat). 
     
     
         16 . The bispecific or multispecific antibody of any of  claims 12  to  15 , wherein the Fc domain comprises a modification promoting the association of the first and second subunit of the Fc domain. 
     
     
         17 . The bispecific or multispecific antibody of any of  claims 12  to  16 , wherein the first subunit of the Fc domain comprises knobs and the second subunit of the Fe domain comprises holes according to the knobs-into-holes technology. 
     
     
         18 . The bispecific or multispecific antibody of any of  claims 12  to  17 , wherein the first subunit of the Fc domain comprises the amino acid substitutions S354C and T366W (numbering according to EU index according to Kabat) and the second subunit of the Fc domain comprises the amino acid substitutions Y349C, T366S and Y407V (numbering according to EU index according to Kabat). 
     
     
         19 . The bispecific or multispecific antibody of any of  claims 12  to  17 , wherein the Fc domain is fused to the C-terminus of the aVH domain, wherein the fusion comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 99, SEQ ID NO: 101, SEQ ID NO: 103, SEQ ID NO: 105, SEQ ID NO: 107, SEQ ID NO: 109, SEQ ID NO: 111, SEQ ID NO: 113, SEQ ID NO: 115, SEQ ID NO: 117, SEQ ID NO: 117; particularly from the group consisting of SEQ ID NO: 105, SEQ ID NO: 107, SEQ ID NO: 109, SEQ ID NO: 111. 
     
     
         20 . The bispecific or multispecific antibody of any of  claims 12  to  18 , wherein the variable domains VL and VH of the Fab fragment comprising the antigen-binding site that binds to PD1 are replaced by each other. 
     
     
         21 . The bispecific or multispecific antibody of any of  claims 12  to  19 , wherein in the Fab fragment in the constant domain CL the amino acid at position 124 is substituted independently by lysine (K), arginine (R) or histidine (H) (numbering according to EU index according to Kabat), and in the constant domain CH1 the amino acids at positions 147 and 213 are substituted independently by glutamic acid (E) or aspartic acid (D) (numbering according to EU index according to Kabat). 
     
     
         22 . The bispecific or multispecific antibody of any of  claims 1  to  21 , comprising
 (a) a first heavy chain comprising an amino acid sequence with at least 95% sequence identity to the sequence of SEQ ID NO: 192, a first light chain comprising an amino acid sequence with at least 95% sequence identity to the sequence of SEQ ID NO: 193, a second heavy chain comprising an amino acid sequence with at least 95% sequence identity to the sequence selected from the group consisting of SEQ ID NO: 99, SEQ ID NO: 101, SEQ ID NO: 103, SEQ ID NO: 105, SEQ ID NO: 107, SEQ ID NO: 109, SEQ ID NO: 111, SEQ ID NO: 113, SEQ ID NO: 115, SEQ ID NO: 117, SEQ ID NO: 117; particularly from the group consisting of SEQ ID NO: 105, SEQ ID NO: 107, SEQ ID NO: 109, SEQ ID NO: 111. 
 
     
     
         23 . The bispecific or multispecific antibody of any of  claims 1  to  21 , comprising
 (a) a heavy chain comprising an amino acid sequence with at least 95% sequence identity to the sequence of SEQ ID NO: 143, or a light chain comprising an amino acid sequence with at least 95% sequence identity to the sequence of SEQ ID NO: 145, and 
 b) a second heavy chain comprising an amino acid sequence with at least 95% sequence identity to the sequence selected from the group consisting of SEQ ID NO: 99, SEQ ID NO: 101, SEQ ID NO: 103, SEQ ID NO: 105, SEQ ID NO: 107, SEQ ID NO: 109, SEQ ID NO: 111, SEQ ID NO: 113, SEQ ID NO: 115, SEQ ID NO: 117, SEQ ID NO: 117; particularly from the group consisting of SEQ ID NO: 105, SEQ ID NO: 107, SEQ ID NO: 109, SEQ ID NO: 111. 
 
     
     
         24 . A polynucleotide encoding for the bispecific or multispecific antibody of any of  claims 1  to  23 . 
     
     
         25 . A vector, particularly an expression vector, comprising the polynucleotide of  claim 24   
     
     
         26 . A host cell, particularly a eukaryotic or prokaryotic host cell, comprising the polynucleotide according to  claim 24  or the vector according to  claim 25 . 
     
     
         27 . A method for producing the bispecific antibody of any of  claims 1  to  23 , comprising the steps of
 (a) transforming a host cell with at least one vector comprising polynucleotides encoding said bispecific or multispecific antibody, 
 (b) culturing the host cell under conditions suitable for the expression of the bispecific or multispecific antibody, and optionally 
 (c) recovering the bispecific or multispecific antibody from the culture, particularly the host cells. 
 
     
     
         28 . A pharmaceutical composition comprising the bispecific or multispecific antibody of any of  claims 1  to  23  and at least one pharmaceutically acceptable excipient. 
     
     
         29 . The bispecific or multispecific antibody of any of  claims 1  to  23  or the pharmaceutical composition according to  claim 28  for use as a medicament. 
     
     
         30 . The bispecific or multispecific antibody of any one of  claims 1  to  23  or the pharmaceutical composition according to  claim 28  for use
 i) in the modulation of immune responses, such as restoring T cell activity, 
 ii) in stimulating an immune response or function, 
 iii) in the treatment of infections, 
 iv) in the treatment of cancer, 
 v) in delaying progression of cancer, 
 vi) in prolonging the survival of a patient suffering from cancer. 
 
     
     
         31 . The bispecific or multispecific antibody of any one of  claims 1  to  23  or the pharmaceutical composition according to  claim 28  for use in the prevention or treatment of cancer. 
     
     
         32 . The bispecific or multispecific antibody of any one of  claims 1  to  23  or the pharmaceutical composition according to  claim 28  for use in the treatment of a chronic viral infection. 
     
     
         33 . The bispecific or multispecific antibody of any one of  claims 1  to  23  or the pharmaceutical composition according to  claim 28  for use in the prevention or treatment of cancer, wherein the bispecific or multispecific antibody is administered in combination with a chemotherapeutic agent, radiation and/or other agents for use in cancer immunotherapy. 
     
     
         34 . A method of inhibiting the growth of tumor cells in an individual comprising administering to the individual an effective amount of the bispecific or multispecific antibody according to any one of  claims 1  to  23  to inhibit the growth of the tumor cells. 
     
     
         35 . The invention as described hereinbefore.